JoVE Journal > Neuroscience
Summary
Abstract
Introduction
Protocol
Ergebnisse
Discussion
Offenlegungen
Acknowledgements
Materials
Referenzen
Please note that all translations are automatically generated. Click here for the English version.
Neurowissenschaften

Chronic Intermittent Ethanol Vapor Exposure Paired with Two-Bottle Choice to Model Alcohol Use Disorder

Published: June 23, 2023
doi:
10.3791/65320
, , , ,
1Department of Basic Medical Sciences, College of Veterinary Medicine,Purdue University, 2The Scripps Research Institute, 3Purdue Institute for Integrative Neuroscience, 4Weldon School of Biomedical Engineering,Purdue University, 5Purdue Institute of Inflammation, Immunology, and Infectious Disease

Summary

Presented here is a protocol for the 2BC/CIE model of alcohol dependence in mice to study alcohol use disorder.

Abstract

Alcohol use disorder (AUD) is a chronic alcohol-related disorder that typically presents as uncontrolled drinking and preoccupation with alcohol. A key component of AUD research is using translationally relevant preclinical models. Over the past several decades, a variety of animal models have been used to study AUD. One prominent model of AUD is the chronic intermittent ethanol vapor exposure (CIE) model, which is a well-established approach for inducing alcohol dependence in rodents through repeated cycles of ethanol exposure via inhalation. To model AUD in mice, the CIE exposure is paired with a voluntary two-bottle choice (2BC) of alcohol drinking and water to measure the escalation of alcohol drinking. The 2BC/CIE procedure involves alternating weeks of 2BC drinking and CIE, which repeat until the escalation of alcohol drinking is achieved. In the present study, we outline the procedures for performing 2BC/CIE, including the daily use of the CIE vapor chamber, and provide an example of escalated alcohol drinking in C57BL/6J mice using this approach.

Introduction

Alcohol use disorder (AUD), which involves chronic excessive alcohol consumption, is one of the most common psychiatric disorders and is a global problem. AUD symptoms involve repeated cycles of intoxication, withdrawal, and cravings and are characterized by the constant consumption of alcohol without regard for the social, occupational, and health consequences1,2,3,4,5,6,7. Alcohol use disorder often occurs in conjunction with other pervasive, persistent, and impairing mental disorders8, such as ADHD9, anxiety10, or depression11 and is responsible for approximately 88,000 deaths annually in the United States alone2. Excessive or frequent alcohol use can affect a person's work status and social relationships12 and may lead to increased violence13. Physically, acute withdrawal from alcohol can result in anxiety, agitation, tremor, excessive sweating, altered consciousness, and hallucinations14,15. Furthermore, people may feel withdrawal symptoms when cutting down or stopping drinking and become irritable or cranky16. Additionally, chronic alcohol consumption can cause memory loss17 and can result in thiamine deficiency, also known as Wernicke-Korsakoff syndrome (WKS), which contributes significantly to alcohol-induced dementia18.

To further advance AUD research, it is necessary to have translationally relevant animal models of the disease. The most common model of AUD in rodents is chronic intermittent ethanol vapor exposure (CIE), which is a well-established approach for inducing alcohol dependence through repeated inhalation of alcohol vapor4,19,20,21,22,23,24,25,26,27,28,29,30. Rodent CIE procedures induce withdrawal symptoms such as handling-induced convulsions31, hyperexcitability, irritability-like behavior, anxiety-like behavior, and sleep disorders and result in an escalation of alcohol drinking22,32,33,34,35, thus meaning the CIE model has translational validity to human AUD.

In rats, the CIE model often involves the operant self-administration of alcohol to measure the escalation of intake36,37,38, whereas the mouse model involves CIE and two-bottle choice (2BC) drinking39,40. Preclinical models of alcohol dependence have consistently shown that animals increase their ethanol intake after chronic ethanol vapor exposure23,41,42,43. In mice specifically, repeated cycles of CIE have been shown to escalate voluntary ethanol intake3,21,44,45,46. Overall, prior studies demonstrate that the CIE model is sufficient to increase ethanol consumption and model AUD in rodents.

This study aims to highlight the CIE method for studying AUD and, more specifically, focus on the 2BC/CIE mouse model. We go through a detailed process of the steps necessary for performing 2BC/CIE and present an example of the escalation of alcohol drinking after CIE.

Protocol

All procedures were approved by the Purdue University Animal Care and Use Committee. The present study used 8 week old C57BL/6J mice. The CIE group had 5 mice (3 male, 2 female), and the Air group had 10 mice (5 male and 5 female). The animals were obtained from a commercial source (see Table of Materials) and were group-housed on a 12 h light-dark cycle with access to food. The body weights of the mice were measured once per week throughout the experiment. 1. General ex…

Representative Results

In the present representative study, ethanol intake (g/kg) during 2BC is reported during baseline drinking and after weeks of CIE (post vapor). Briefly, as described in the protocol, during 2BC, the mice had access to two bottles: one containing water and the other containing 15% (w/v) ethanol. After the baseline intakes were determined, the subjects were split and evenly assigned to the CIE or Air group. The initial baseline ethanol intake during the 3 week period stabilized at 2.00 g/kg ± 0.21 g/kg (n = 15) before…

Discussion

Alcohol use disorder represents a global public health problem with high prevalence and cost to society52. To study AUD in preclinical animal models, a common method in mice is 2BC/CIE20,34,39,40,47,53,54,55. Here, this established m…

Offenlegungen

The authors have nothing to disclose.

Acknowledgements

This work was supported by National Institutes of Health (NIH) grants AA027301 and AA029985.

Materials

500 Eppendorf Tubes Eppendorf L203896J
95% ethanol Decon laboratories 2816
Analox machine Analox Instruments Analox-AM1
Animal Weighing Scale Kent Scientific SCL-4000
Binder Clips Office Depot 560394
C57BL/6J mice The Jackson Laboratory 000664
Centrifuge Eppendorf 5418R
Chronic intermitted vapor chamber La Jolla Alcohol Research Inc Custom made materials
Heparin/EDTA Sagent Pharmaceuticals TS/DRUGS/2/2015
Mouse bedding Bed-o’Cobs 8B fill with 1/8" deep
Mouse drinking bottle Custom made materials
Pyrazole Sigma-Aldrich bccc6397
Teklad global 18% protein (mouse food) Teklad global Envigo 2018
DOWNLOAD MATERIALS LIST

Referenzen

  1. Koob, G. F., Volkow, N. D. Neurobiology of addiction: A neurocircuitry analysis. Lancet Psychiatry. 3 (8), 760-773 (2016).
  2. Witkiewitz, K., Litten, R. Z., Leggio, L. Advances in the science and treatment of alcohol use disorder. Science Advances. 5 (9), 4043 (2019).
  3. Becker, H. C. Alcohol dependence, withdrawal, and relapse. Alcohol Research and Health. 31 (4), 348-361 (2008).
  4. Becker, H. C. Effects of alcohol dependence and withdrawal on stress responsiveness and alcohol consumption. Alcohol Research. 34 (4), 448-458 (2012).
  5. Erickson, E. K., Grantham, E. K., Warden, A. S., Harris, R. A. Neuroimmune signaling in alcohol use disorder. Pharmacology, Biochemistry, and Behavior. 177, 34-60 (2019).
  6. Aalsma, M., et al. Adolescent suicide assessment and management in primary care. BMC Pediatrics. 22 (1), 389 (2022).
  7. Grant, B. F., et al. Epidemiology of DSM-5 alcohol use disorder: Results from the National Epidemiologic Survey on Alcohol and Related Conditions III. JAMA Psychiatry. 72 (8), 757-766 (2015).
  8. Helle, A. C., Watts, A. L., Trull, T. J., Sher, K. J. Alcohol use disorder and antisocial and borderline personality disorders. Alcohol Research. 40 (1), (2019).
  9. Luderer, M., Ramos Quiroga, J. A., Faraone, S. V., Zhang James, Y., Reif, A. Alcohol use disorders and ADHD. Neuroscience and Biobehavioral Reviews. 128, 648-660 (2021).
  10. Ribadier, A., Varescon, I. Anxiety and depression in alcohol use disorder individuals: The role of personality and coping strategies. Substance Use and Misuse. 54 (9), 1475-1484 (2019).
  11. McHugh, R. K., Weiss, R. D. Alcohol use disorder and depressive disorders. Alcohol Research. 40 (1), (2019).
  12. Le Berre, A. P. Emotional processing and social cognition in alcohol use disorder. Neuropsychology. 33 (6), 808-821 (2019).
  13. Sontate, K. V., et al. Alcohol, aggression, and violence: From public health to neuroscience. Frontiers in Psychology. 12, 699726 (2021).
  14. Prescrire International. Alcohol withdrawal syndrome: how to predict, prevent, diagnose and treat it. Prescrire International. 16 (87), 24-31 (2007).
  15. Mendelson, J. H. The effect of Alcohol on the nervous system. Alcohol Health and Research World. 19 (1), 28-29 (1995).
  16. Murray, T. D., Berger, A. Alcohol withdrawal. Virginia Medical Quarterly. 124 (3), 184-187 (1997).
  17. Fama, R., et al. Memory impairment in alcohol use disorder is associated with regional frontal brain volumes. Drug and Alcohol Dependence. 228, 109058 (2021).
  18. Kopelman, M. D. The Korsakoff syndrome. British Journal of Psychiatry. 166 (2), 154-173 (1995).
  19. Griffin, W. C. Alcohol dependence and free-choice drinking in mice. Alcohol. 48 (3), 287-293 (2014).
  20. Griffin, W. C., Lopez, M. F., Yanke, A. B., Middaugh, L. D., Becker, H. C. Repeated cycles of chronic intermittent ethanol exposure in mice increases voluntary ethanol drinking and ethanol concentrations in the nucleus accumbens. Psychopharmacology. 201 (4), 569-580 (2009).
  21. Kimbrough, A., et al. CRF(1) receptor-dependent increases in irritability-like behavior during abstinence from chronic intermittent ethanol vapor exposure. Alcoholism: Clinical and Experimental Research. 41 (11), 1886-1895 (2017).
  22. Kimbrough, A., et al. Brain-wide functional architecture remodeling by alcohol dependence and abstinence. Proceedings of the National Academy of Sciences of the United States of America. 117 (4), 2149-2159 (2020).
  23. O’Dell, L. E., Roberts, A. J., Smith, R. T., Koob, G. F. Enhanced alcohol self-administration after intermittent versus continuous alcohol vapor exposure. Alcoholism: Clinical and Experimental Research. 28 (11), 1676-1682 (2004).
  24. Pleil, K. E., et al. Effects of chronic ethanol exposure on neuronal function in the prefrontal cortex and extended amygdala. Neuropharmacology. 99, 735-749 (2015).
  25. Warden, A. S., et al. Microglia control escalation of drinking in alcohol-dependent mice: Genomic and synaptic drivers. Biological Psychiatry. 88 (12), 910-921 (2020).
  26. Gilpin, N. W., Koob, G. F. Neurobiology of alcohol dependence: Focus on motivational mechanisms. Alcohol Research and Health. 31 (3), 185-195 (2008).
  27. Kreifeldt, M., et al. Central amygdala corticotropin-releasing factor neurons promote hyponeophagia but do not control alcohol drinking in mice. Molecular Psychiatry. 27 (5), 2502-2513 (2022).
  28. Sidhu, H., Kreifeldt, M., Contet, C. Affective disturbances during withdrawal from chronic intermittent ethanol inhalation in C57BL/6J and DBA/2J male mice. Alcoholism: Clinical and Experimental Research. 42 (7), 1281-1290 (2018).
  29. Kononoff, J., et al. Systemic and intra-habenular activation of the orphan G protein-coupled receptor GPR139 decreases compulsive-like alcohol drinking and hyperalgesia in alcohol-dependent rats. eNeuro. 5 (3), (2018).
  30. Avegno, E. M., et al. Central amygdala circuits mediate hyperalgesia in alcohol-dependent rats. Journal of Neuroscience. 38 (36), 7761-7773 (2018).
  31. de Guglielmo, G., et al. Recruitment of a neuronal ensemble in the central nucleus of the amygdala is required for alcohol dependence. Journal of Neuroscience. 36 (36), 9446-9453 (2016).
  32. Liang, J., et al. Chronic intermittent ethanol-induced switch of ethanol actions from extrasynaptic to synaptic hippocampal GABAA receptors. Journal of Neuroscience. 26 (6), 1749-1758 (2006).
  33. Becker, H. C. Animal models of alcohol withdrawal. Alcohol Research and Health. 24 (2), 105-113 (2000).
  34. Kimbrough, A., Kim, S., Cole, M., Brennan, M., George, O. Intermittent access to ethanol drinking facilitates the transition to excessive drinking after chronic intermittent ethanol vapor exposure. Alcoholism: Clinical and Experimental Research. 41 (8), 1502-1509 (2017).
  35. Koob, G. F., Colrain, I. M. Alcohol use disorder and sleep disturbances: A feed-forward allostatic framework. Neuropsychopharmacology. 45 (1), 141-165 (2020).
  36. Risher, M. L., et al. Long-term effects of chronic intermittent ethanol exposure in adolescent and adult rats: Radial-arm maze performance and operant food reinforced responding. PLoS One. 8 (5), e62940 (2013).
  37. Van Skike, C. E., Novier, A., Diaz-Granados, J. L., Matthews, D. B. The effect of chronic intermittent ethanol exposure on spatial memory in adolescent rats: The dissociation of metabolic and cognitive tolerances. Brain Research. 1453, 34-39 (2012).
  38. Ho, A. M., et al. Chronic intermittent ethanol exposure alters behavioral flexibility in aged rats compared to adult rats and modifies protein and protein pathways related to Alzheimer’s disease. ACS Omega. 7 (50), 46260-46276 (2022).
  39. Piggott, V. M., Lloyd, S. C., Perrine, S. A., Conti, A. C. Chronic intermittent ethanol exposure increases ethanol consumption following traumatic stress exposure in mice. Frontiers in Behavioral Neuroscience. 14, 114 (2020).
  40. Quijano Carde, N. A., De Biasi, M. Behavioral characterization of withdrawal following chronic voluntary ethanol consumption via intermittent two-bottle choice points to different susceptibility categories. Alcoholism: Clinical and Experimental Research. 46 (4), 614-627 (2022).
  41. Rimondini, R., Arlinde, C., Sommer, W., Heilig, M. Long-lasting increase in voluntary ethanol consumption and transcriptional regulation in the rat brain after intermittent exposure to alcohol. FASEB Journal. 16 (1), 27-35 (2002).
  42. Becker, H. C. Influence of stress associated with chronic alcohol exposure on drinking. Neuropharmacology. 122, 115-126 (2017).
  43. Zhao, Y., Weiss, F., Zorrilla, E. P. Remission and resurgence of anxiety-like behavior across protracted withdrawal stages in ethanol-dependent rats. Alcoholism: Clinical and Experimental Research. 31 (9), 1505-1515 (2007).
  44. Becker, H. C., Lopez, M. F. Increased ethanol drinking after repeated chronic ethanol exposure and withdrawal experience in C57BL/6 mice. Alcoholism: Clinical and Experimental Research. 28 (12), 1829-1838 (2004).
  45. Kreifeldt, M., Le, D., Treistman, S. N., Koob, G. F., Contet, C. BK channel beta1 and beta4 auxiliary subunits exert opposite influences on escalated ethanol drinking in dependent mice. Frontiers in Integrative Neuroscience. 7, 105 (2013).
  46. Pringuey, D., et al. Comorbidity of affective disorders and alcohol use disorder. Encephale. 40, S3-S7 (2014).
  47. Okhuarobo, A., et al. A novel mouse model for vulnerability to alcohol dependence induced by early-life adversity. Neurobiology of Stress. 13, 100269 (2020).
  48. Dilley, J. E., Nicholson, E. R., Fischer, S. M., Zimmer, R., Froehlich, J. C. Alcohol drinking and blood alcohol concentration revisited. Alcoholism: Clinical and Experimental Research. 42 (2), 260-269 (2018).
  49. Feierman, D. E., Cederbaum, A. I. Oxidation of the alcohol dehydrogenase inhibitor pyrazole to 4-hydroxypyrazole by microsomes. Effect of cytochrome P-450 inducing agents. Drug Metabolism and Disposition. 15 (5), 634-639 (1987).
  50. Parasuraman, S., Raveendran, R., Kesavan, R. Blood sample collection in small laboratory animals. Journal of Pharmacology and Pharmacotherapeutics. 1 (2), 87-93 (2010).
  51. Armstrong, R. A. When to use the Bonferroni correction. Ophthalmic and Physiological Optics. 34 (5), 502-508 (2014).
  52. Rehm, J., et al. Alcohol use disorders in primary health care: What do we know and where do we go. Alcohol and Alcoholism. 51 (4), 422-427 (2016).
  53. Gorini, G., Roberts, A. J., Mayfield, R. D. Neurobiological signatures of alcohol dependence revealed by protein profiling. PLoS One. 8 (12), e82656 (2013).
  54. Frie, J. A., Khokhar, J. Y. An open source automated two-bottle choice test apparatus for rats. HardwareX. 5, 00061 (2019).
  55. Smith, R. J., et al. Dynamic c-Fos changes in mouse brain during acute and protracted withdrawal from chronic intermittent ethanol exposure and relapse drinking. Addiction Biology. 25 (6), e12804 (2020).
  56. Sachdeva, A., Choudhary, M., Chandra, M. Alcohol withdrawal syndrome: Benzodiazepines and beyond. Journal of Clinical and Diagnostic Research. 9 (9), (2015).
  57. Huitron-Resendiz, S., et al. Effects of withdrawal from chronic intermittent ethanol exposure on sleep characteristics of female and male mice. Alcoholism: Clinical and Experimental Research. 42 (3), 540-550 (2018).
  58. Gatch, M. B., Lal, H. Effects of ethanol and ethanol withdrawal on nociception in rats. Alcoholism: Clinical and Experimental Research. 23 (2), 328-333 (1999).
  59. Saitz, R. Introduction to alcohol withdrawal. Alcohol Health and Research World. 22 (1), 5-12 (1998).
  60. Emmerson, R. Y., Dustman, R. E., Shearer, D. E., Chamberlin, H. M. EEG, visually evoked and event related potentials in young abstinent alcoholics. Alcohol. 4 (4), 241-248 (1987).
  61. Gauvin, D. V., Youngblood, B. D., Holloway, F. A. The discriminative stimulus properties of acute ethanol withdrawal (hangover) in rats. Alcoholism: Clinical and Experimental Research. 16 (2), 336-341 (1992).
  62. George, O., Le Moal, M., Koob, G. F. Allostasis and addiction: Role of the dopamine and corticotropin-releasing factor systems. Physiology and Behavior. 106 (1), 58-64 (2012).
  63. Koob, G. F., Le Moal, M. Addiction and the brain antireward system. Annual Review of Psychology. 59, 29-53 (2008).
  64. Koob, G. F., Le Moal, M. Drug addiction, dysregulation of reward, and allostasis. Neuropsychopharmacology. 24 (2), 97-129 (2001).
  65. Nieto, S. J., Grodin, E. N., Aguirre, C. G., Izquierdo, A., Ray, L. A. Translational opportunities in animal and human models to study alcohol use disorder. Translational Psychiatry. 11 (1), 496 (2021).
  66. Kuhns, L., Kroon, E., Lesscher, H., Mies, G., Cousijn, J. Age-related differences in the effect of chronic alcohol on cognition and the brain: A systematic review. Translational Psychiatry. 12 (1), 345 (2022).
  67. Becker, H. C., Mulholland, P. J. Neurochemical mechanisms of alcohol withdrawal. Handbook of Clinical Neurology. 125, 133-156 (2014).
  68. Chen, M. M., et al. Intoxication by intraperitoneal injection or oral gavage equally potentiates postburn organ damage and inflammation. Mediators of Inflammation. , 971481 (2013).
  69. Livy, D. J., Parnell, S. E., West, J. R. Blood ethanol concentration profiles: A comparison between rats and mice. Alcohol. 29 (3), 165-171 (2003).
  70. Jin, L., Batra, S., Jeyaseelan, S. Diminished neutrophil extracellular trap (NET) formation is a novel innate immune deficiency induced by acute ethanol exposure in polymicrobial sepsis, which can be rescued by CXCL1. PLoS Pathogens. 13 (9), e1006637 (2017).
  71. Crabbe, J. C., Harris, R. A., Koob, G. F. Preclinical studies of alcohol binge drinking. Annals of the New York Academy of Sciences. 1216, 24-40 (2011).
  72. Blomstrand, R., et al. Pyrazoles as inhibitors of alcohol oxidation and as important tools in alcohol research: An approach to therapy against methanol poisoning. Proceedings of the National Academy of Sciences of the United States of America. 76 (7), 3499-3503 (1979).
  73. Domi, A., Stopponi, S., Domi, E., Ciccocioppo, R., Cannella, N. Sub-dimensions of alcohol use disorder in alcohol preferring and non-preferring rats, a comparative study. Frontiers in Behavioral Neuroscience. 13, 3 (2019).
  74. de Guglielmo, G., et al. Voluntary and forced exposure to ethanol vapor produces similar escalation of alcohol drinking but differential recruitment of brain regions related to stress, habit, and reward in male rats. Neuropharmacology. 222, 109309 (2023).
  75. Haggerty, D. L., et al. The role of anterior insular cortex inputs to dorsolateral striatum in binge alcohol drinking. Elife. 11, 77411 (2022).
  76. Slivicki, R. A., et al. Oral oxycodone self-administration leads to features of opioid misuse in male and female mice. Addiction Biology. 28 (1), e13253 (2023).
  77. Godynyuk, E., Bluitt, M. N., Tooley, J. R., Kravitz, A. V., Creed, M. C. An open-source, automated home-cage sipper device for monitoring liquid ingestive behavior in rodents. eNeuro. 6 (5), (2019).
  78. Simpson, S., et al. The hidden brain: Uncovering previously overlooked brain regions by employing novel preclinical unbiased network approaches. Frontiers in Systems Neuroscience. 15, 595507 (2021).
  79. Smith, L. C., Kimbrough, A. Leveraging neural networks in preclinical alcohol research. Brain Sciences. 10 (9), 578 (2020).
  80. Hwa, L., Besheer, J., Kash, T. Glutamate plasticity woven through the progression to alcohol use disorder: A multi-circuit perspective. F1000Research. 6, 298 (2017).

Tags

Alcohol Use DisorderChronic Intermittent Ethanol Vapor ExposureTwo-bottle ChoiceRodent ModelAlcohol DependenceVoluntary Alcohol DrinkingEscalation Of Alcohol IntakeC57BL/6J Mice
This article has been published
Video Coming Soon
Keep me updated:

.

PDF
DOI
DOWNLOAD MATERIALS LIST
Diesen Artikel zitieren
Xiao, T., Chen, Y., Boisvert, A., Cole, M., Kimbrough, A. Chronic Intermittent Ethanol Vapor Exposure Paired with Two-Bottle Choice to Model Alcohol Use Disorder. J. Vis. Exp. (196), e65320, doi:10.3791/65320 (2023).
Zitat herunterladen
Nachdrucke und Berechtigungen

View Video

✖

To prove you're not a robot, please enter the text in the image below

CAPTCHA Image
Play CAPTCHA Audio
Refresh Image